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Purification of gonad-stimulating substance (GSS) and its secretion mechanism during breeding season

繁殖期性腺刺激物质（GSS）的纯化及其分泌机制

基本信息

批准号：
13640673
负责人：
MITA Masatoshi
金额：
$ 1.79万
依托单位：
Teikyo University (2002)Teikyo Junior College (2001)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

In starfish, oocyte maturation and spawning are first triggered by a peptide hormone (gonad-stimulating substance, GSS) present in nervous tissue. GSS was purified from radial nerves of starfish Asterina pectinifera. Although a molecular weight of A. pectinifera GSS was estimated to about 4,700 using a mass-spectrometry, the quantity was not enough to determine the amino acid sequence. This study also examined organization of neural networks in ovaries and testes of the starfish, Asterina pectinifera by employing an antibody that recognizes molecules belonging to SALMFamide family of echinoderm neuropeptides. Following incubation with a SALMFamide 1 antibody primary antibody, and subsequently with rhodamine-conjugated second antibody identified the network structures were observed in both ovaries and testes. The networks were restricted mainly to their surface and little evidence of immunoreactivity was seen inside their basement membranes. The result suggests that neural networks distributed in gonads maybe involved in the regulation of gametogenesis and gamete shedding in starfish.Furthermore, using degenerate primers derived from consensus sequences of the memmalian glycoprotein hormone (LH, FSH and TSH) receptors, we cloned cDNAs (apGPHR) from ovaries of.the starfish Asterina pectinifera. However, a partially purified gonad-stimulating substance (GSS) failed to increase cyclic AMP levels in COS-7 cells expressing apGPHR. This suggests that a new neural petide other than GSS is a ligand for the apGPHR in starfish.

在海星中，卵母细胞成熟和产卵首先由神经组织中存在的肽激素（性腺刺激物质，GSS）触发。GSS是从海星（Asterina pectinifera）的桡神经中分离纯化的。虽然A.使用质谱法估计pectinifera GSS为约4，700，该量不足以确定氨基酸序列。本研究还研究了海星，海星的卵巢和睾丸中的神经网络的组织，采用抗体识别分子属于SALMFamide家庭棘皮动物神经肽。与SALMF酰胺1抗体一抗孵育后，随后与罗丹明缀合的二抗孵育，在卵巢和睾丸中观察到网络结构。这些网络主要局限于其表面，在其基底膜内几乎看不到免疫反应性的证据。本研究还利用膜糖蛋白激素（LH、FSH和TSH）受体的保守序列设计简并引物，从栉孔海星（Asterina pectinifera）卵巢中克隆了apGPHR基因。然而，部分纯化的性腺刺激物质（GSS）未能增加表达apGPHR的COS-7细胞中的环AMP水平。这表明，一个新的神经肽以外的GSS是海星的apGPHR的配体。